A novel approach to quantitative characterisation of key volatile aroma compounds in strawberry (Fragaria × ananassa) using stable isotope dilution assay and HS-SPME-GC-MS for flavour-driven breeding.

Strawberry flavour is determined by a complex mix of non-volatile compounds like sugars and organic acids, along with volatile aroma compounds that significantly impact fruit quality. With strawberry production rapidly expanding worldwide, strawberry breeding programs are focusing on improving and diversifying berry quality and flavour to meet consumer demand. This study reports the development and evaluation of an accurate and precise analytical method for the simultaneous quantification of thirteen key volatile aroma compounds using stable isotope dilution assay combined with headspace solid-phase microextraction and gas chromatography-mass spectrometry. Calibration functions of the matrix-matched validated method showed high accuracy with R2 values between 0.9624 and 0.9997. The method was applied to identify the key aroma volatile compounds produced by ten strawberry cultivars and breeding accessions. This study provides an accurate and precise analytical framework that supports breeding for improved sensory aroma profiles, ultimately contributing to the development of superior strawberry varieties tailored to consumer preferences.